Infected Animals Research Articles

Retrospective Evaluation of Pediatric Patients Hospitalized with Brucellosis: Single Center Study in Istanbul

Aim: Brucellosis is a zoonotic disease and mainly develops as a result of consuming products of infected animals such as cattle, sheep and pigs. Material-Methods: Pediatric patients under the age of 18 who were hospitalized with a diagnosis of brucellosis in a third-level city hospital were included in the study. Serology titer ≥1:160 and/or Brucella spp in blood culture growth was determined as the diagnostic criterion for the diagnosis of brucellosis. Results: Of the 35 pediatric patients diagnosed with brucellosis, 22 were boys and 13 were girls, and the average age was determined as 154.2 ± 53.1 months (25-214). Hospital admission complaints were determined in order of frequency as follows; joint pain (51.4%), fever (31.4%), headache (17.1%) and abdominal pain (5.2%). Eleven of the patients (31.4%) had polyarthralgia and 7 (20%) had monoarthralgia. The most common physical examination findings were hepatomegaly (48.6%) and splenomegaly (40%), respectively. Joint involvement was detected in 17.1% of the patients. Sacroiliac joint involvement was reported in 3 (8.6%) patients, ankle joint involvement in 2 (5.7%) patients, and knee joint involvement in 1 (2.9%) patient. Anemia (60%) was the most common hematological finding. The average hemoglobin value was determined as 12.5±1.44 (10.2-15.9) g/dl. When patients are compared in terms of joint involvement; neutrophil count and treatment duration were found to be statistically higher in patients with joint involvement (p<0.05). Conclusion: Brucellosis is a zoonotic disease that is endemic in our country, affects all age groups, and can cause complications and relapses, and continues to be a serious public health problem for our country.

Epidemiology and antibiotic resistance of staphylococci on commercial pig farms in Cape Town, South Africa

Staphylococci are responsible for a wide range of infections in animals. The most common species infecting animals include Staphylococcus aureus, Staphylococcus epidermidis and Staphylococcus intermedius. Recent increases in antibiotic use and antibiotic resistance in animals highlight the need to understand the potential role of commercial livestock as a reservoir of staphylococci and antibiotic resistance genes. Nasal swabs were collected from 143 apparently healthy pigs and 21 pig farm workers, and 45 environmental swabs of feed and water troughs, from two commercial pig farms in the Western Cape, South Africa. Staphylococci were isolated, identified using mass-spectrometry, and antimicrobial susceptibility testing and Illumina whole genome sequencing were performed. One hundred and eighty-five (185) Staphylococcus spp. isolates were obtained, with Mammalicoccus sciuri (n = 57; 31%) being the most common, followed by S. hyicus (n = 40; 22%) and S. aureus (n = 29; 16%). S. epidermidis was predominantly identified in the farm workers (n = 18; 86%). Tetracycline resistance was observed across all species, with rates ranging from 67 to 100%. Majority of M. sciuri isolates (n = 40; 70%) were methicillin resistant, with 78% (n = 31) harbouring mecA. M. sciuri isolates had genes/elements which were associated with SCCmec_type_III (3A) and SCCmec_type_VIII(4A) and were mostly observed in ST61 strains. ST239 strains were associated with SCCmec_type_III(3A). High rates of tetracycline resistance were identified among staphylococci in the pig farms in Western Cape, South Africa. This highlights the need for policy makers to regulate the use of this antibiotic in pig farming.

Genomic analysis of two penicillin- and rifampin-resistant Corynebacterium rouxii strains isolated from cutaneous infections in dogs

Although diphtheria is a vaccine-preventable disease, numerous cases are still reported around the world, as well as outbreaks in countries, including European ones. Species of the Corynebacterium diphtheriae complex are potentially toxigenic and, therefore, must be considered given the possible consequences, such as the circulation of clones and transmission of antimicrobial resistance and virulence genes. Recently, Corynebacterium rouxii was characterized and included among the valid species of the complex. Therefore, two cases of C. rouxii infection arising from infections in domestic animals are presented here. We provide molecular characterization, phylogenetic analyses, genome sequencing, and CRISPR-Cas analyses to contribute to a better understanding of the molecular bases, pathogenesis, and epidemiological monitoring of this species, which is still little studied. We confirmed its taxonomic position with genome sequencing and in silico analysis and identified the ST-918 for both strains. The clinical isolates were sensitive resistance to benzylpenicillin and rifampin. Antimicrobial resistance genes, including tetB, rpoB2, and rbpA genes, were predicted. The bla and ampC genes were not found. Several virulence factors were also detected, including adhesion, iron uptake systems, gene regulation (dtxR), and post-translational modification (MdbA). Finally, one prophage and the Type I-E CRISPR-Cas system were identified.

Monkeypox (MPOX): Unmasking the Lesser-known Viral Threat

Monkeypox (Mpox) is caused by a viral infection and has been endemic in several African states after the last outbreak throughout the world in 2003. This disease resurfaced in the year 2022 and has since spread worldwide, even in countries where no cases had been reported in the earlier outbreak. While the world was still recovering from the menace of corona virus disease 2019 this virus; having similar features, started to spread across the world affecting thousands of people. The virus can be transmitted from human to human, and to humans from bites and scratches of infected animals. Common signs and symptoms include rashes and lymphadenopathy, accompanied with fever. Preventive measures for the Mpox virus include: using masks and alcohol-based hand sanitizers, practicing social distancing in crowded areas and practicing safe sexual activities. ACAM2000 and LC16m8 are a few vaccines available for its treatment. Early diagnosis and timely treatment of the disease have helped to curb the incidence and mortality of Mpox. Through this article, we discuss the history of infection, mode of transmission, the genomic data of the virus along with its signs and symptoms and its prevention and treatment using available vaccines. World governments and healthcare providers need to act appropriately in order to combat another such outbreak, as this disease has not been eradicated yet.

Towards Safe African Swine Fever Vaccines: The A137R Gene as a Tool to Reduce Virulence and a Promising Serological DIVA Marker Candidate.

African swine fever (ASF) is an emerging disease caused by the African swine fever virus (ASFV), which is a great threat to the swine industry worldwide. Currently registered vaccines that have demonstrated protection against the homologous ASFV strains are live attenuated vaccines based on recombinant ASFV strains with the deletions of virulence-associated genes. In this study, we evaluated the deletion of the A137R gene in the ASFV virulent Stavropol_01/08 strain isolated in Russia in 2008. Our animal experiment results demonstrated that the deletion of the A137R gene did not lead to the full attenuation of this strain, and increasing the dose of the A137R-deletion mutant during infection led to the death of 87.5% of the infected animals. In this report, we also demonstrated that immunofluorescence (IFA) and Western blotting assays based on the recombinant p11.5 protein can be used to detect antibodies in animals infected with the attenuated ASFV variants of several genotypes/serotypes. Both assays were specific to ASFV p11.5 protein and showed negative results when examining the sera of the non-infected animals or those infected with the A137R-deletion mutant. Therefore, we propose to use the p11.5 protein along with other previously proposed ASFV proteins, such as CD2v, as negative antigenic DIVA markers for an attenuated ASF vaccine.

Molecular and serological screening of coxiellosis in bovines with reproductive disorders in dairy farms

Coxiella burnetii infection is an emerging/re-emerging public health problem affecting several countries worldwide. In India, the disease is mainly underdiagnosed, creating hindrances in its effective control. This study investigated the occurrence of C. burnetii among apparently healthy cattle and cattle with a history of reproductive disorders by both PCR and indirect-ELISA. A total of 731 clinical samples (serum: 531, and vaginal swabs as well as blood: 100 each) from 531 cattle were screened for coxiellosis. The serum, blood, and vaginal swabs each collected from 100 cattle with a history of reproductive disorders were screened using Com1-PCR, Trans-PCR, and indirect-ELISA. Conversely, serum samples obtained from apparently healthy cattle were exclusively screened using indirect ELISA. None of the samples tested could detect C. burnetii in PCR assays, while 13.37 % of serum samples were found to be seropositive in i-ELISA. Seropositivity noted among clinically healthy and those suffering from reproductive disorders were 12.76 % and 16 %, respectively, exhibiting a non-significant difference observed between these two categories. The obtained results suggested that the occurrence of coxiellosis did not differ significantly between clinically healthy animals and those with reproductive disorders; hence, in farms affected with C. burnetii infection, screening healthy and symptomatic animals is crucial to implement appropriate preventive measures.

A Systematic Assessment of Leishmania donovani Infection in Domestic and Wild Animal Reservoir Hosts of Zoonotic Visceral Leishmaniasis in India

Leishmaniasis is a neglected disease with a global spread that affects both domestic and wild animals in addition to people. Leishmania donovani is the suspected anthroponotic cause of visceral leishmaniasis (VL) in India, where it is an endemic disease. The reservoir hosts play a crucial role in the life cycle of the Leishmania parasite. The complicated connection between the pathogen, vector, and reservoir exhibits geographical and temporal diversity. Human-to-human and, to a lesser extent, human-to-animal transmission are the principal mechanisms for the maintenance of anthroponotic diseases. A number of animals were examined for the presence of Leishmania parasites and the findings were reviewed in order to examine the role of animal reservoirs in domestic transmission of cutaneous leishmaniasis in endemic regions of India. The analysis objective was to assess the research conducted on domestic animals’ propensity to spread L. donovani in endemic areas, with a particular emphasis on how proximity and animal density may impact the prevalence of human leishmaniasis. Species of the L. donovani complex have distinct enzootic, zoonotic, and anthroponotic life cycles that depend on the environment. The majority of Leishmania spp. are zoonotic, spreading from non-human mammals to humans. Many nations have leishmaniasis as an endemic disease, and the Indian subcontinent (ISC) has an estimated two to three lakh people who are at risk. This systematic review evaluates the gaps in our understanding of disease transmission that contradict conventional wisdom about the reservoir(s) of visceral leishmaniasis and efforts to manage it on the Indian subcontinent. Fundamental concerns in VL epidemiology and ecology will be clarified by a better understanding of L. donovani infection in domestic animals and its transfer to sandflies. A deliberate, systematic search was conducted on PubMed, Science Direct, and Google Scholar using keywords such as “Leishmania donovani”, “zoonotic visceral leishmaniasis”, and “wild animal reservoir for Leishmania donovani”. A total of 530 potentially relevant references were obtained from these databases, and 507 were not considered due to copy avoidance, irrelevant titles, research publications from nations other than India, or modified compositions. Among the remaining 23 investigations, 20 were rejected, and only 3 were included in the present study. Finally, three research papers with 867 goats, 161 cattle, 106 chickens, 26 sheep, three buffaloes, 406 dogs, and 309 rats were reported. Along with these data, studies across Asian and African countries that are considered VL-endemic areas have been discussed. According to the review, goats are the epidemic’s primary host and possible reservoir in several regions of India. In the endemic regions of the disease, some species of rodents, along with the canines, appear to be maintaining the L. donovani transmission cycle.

Endogenous PGD2 acting on DP2 receptor counter regulates Schistosoma mansoni infection-driven hepatic granulomatous fibrosis.

Identifying new molecular therapies targeted at the severe hepatic fibrosis associated with the granulomatous immune response to Schistosoma mansoni infection is essential to reduce fibrosis-related morbidity/mortality in schistosomiasis. In vitro cell activation studies suggested the lipid molecule prostaglandin D2 (PGD2) as a potential pro-fibrotic candidate in schistosomal context, although corroboratory in vivo evidence is still lacking. Here, to investigate the role of PGD2 and its cognate receptor DP2 in vivo, impairment of PGD2 synthesis by HQL-79 (an inhibitor of the H-PGD synthase) or DP2 receptor inhibition by CAY10471 (a selective DP2 antagonist) were used against the fibrotic response of hepatic eosinophilic granulomas of S. mansoni infection in mice. Although studies have postulated PGD2 as a fibrogenic molecule, HQL-79 and CAY10471 amplified, rather than attenuated, the fibrotic response within schistosome hepatic granulomas. Both pharmacological strategies increased hepatic deposition of collagen fibers - an unexpected outcome accompanied by further elevation of hepatic levels of the pro-fibrotic cytokines TGF-β and IL-13 in infected animals. In contrast, infection-induced enhanced LTC4 synthesis in the schistosomal liver was reduced after HQL-79 and CAY10471 treatments, and therefore, inversely correlated with collagen production in granulomatous livers. Like PGD2-directed maneuvers, antagonism of cysteinyl leukotriene receptors CysLT1 by MK571 also promoted enhancement of TGF-β and IL-13, indicating a key down-regulatory role for endogenous LTC4 in schistosomiasis-induced liver fibrosis. An ample body of data supports the role of S. mansoni-driven DP2-mediated activation of eosinophils as the source of LTC4 during infection, including: (i) HQL-79 and CAY10471 impaired systemic eosinophilia, drastically decreasing eosinophils within peritoneum and hepatic granulomas of infected animals in parallel to a reduction in cysteinyl leukotrienes levels; (ii) peritoneal eosinophils were identified as the only cells producing LTC4 in PGD2-mediated S. mansoni-induced infection; (iii) the magnitude of hepatic granulomatous eosinophilia positively correlates with S. mansoni-elicited hepatic content of cysteinyl leukotrienes, and (iv) isolated eosinophils from S. mansoni-induced hepatic granuloma synthesize LTC4 in vitro in a PGD2/DP2 dependent manner. So, our findings uncover that granulomatous stellate cells-derived PGD2 by activating DP2 receptors on eosinophils does stimulate production of anti-fibrogenic cysLTs, which endogenously down-regulates the hepatic fibrogenic process of S. mansoni granulomatous reaction - an in vivo protective function which demands caution in the future therapeutic attempts in targeting PGD2/DP2 in schistosomiasis.

Interleukin-17A (IL-17A) is involved in antibody specificity to conformational epitopes

The specificity of antibodies (Ab) is essential for the precise recognition of foreign or dangerous molecules. We have shown that mice infected with non-pathogenic Lactate Dehydrogenase Elevating Virus (LDV) inoculated with human growth hormone (hGH) or Ovalbumin (OVA), exhibit modified specificity of anti-hGH or anti-OVA Ab associated with the secretion of IFN-γ, IL-13, and IL-17.Cytokines are directly or indirectly involved in the isotypes, specificity, and affinity of Ab. Accordingly, here we investigated the effect of IL-17 neutralization on Ab specificities to OVA and Diphtheria Toxoid (DTx) in a mouse model of viral infection. Thereby, we employed an anti-cytokine “auto-vaccination” with an OVA/IL-17A complex or a Monoclonal Ab (MAb) anti-IL-17A (MM17/F3).Competitive ELISA assays were used to estimate the quality of the humoral immune response and the amount of Abs to conformational versus linear antigenic determinants. Results indicated that the OVA/IL-17A complex increased Abs levels to conformational epitopes of OVA, while LDV prolonged antibodies for a longer period. Mice treated with MM17F3 MAb showed an increase in Abs to conformational epitopes of OVA. A similar effect, confirmed by a competitive Western-blot assay, was produced by LDV. Moreover, an increased level of IgM, IgG1, and IgG2a was found in infected animals. Similarly, MAb anti-IL-17A treatment increased the proportion of Ab to conformational epitopes of DTx in uninfected mice, while LDV decreased this parameter.In conclusion, our findings demonstrate a correlation between IL-17A neutralization and a change in Ab specificity to OVA or DTx, presenting a novel strategy for obtaining Abs with higher specificity.

Dermatophytosis in Companion Animals in Portugal: A Comprehensive Epidemiological Retrospective Study of 12 Years (2012-2023).

Dermatophytosis, commonly referred to as ringworm, is a common superficial fungal infection in companion animals and humans. Between 2012 and 2023, plucked hair and scraped scale samples from domestic dogs and cats with clinical suspicion of dermatophytosis were collected from 355 veterinary medical centres across mainland Portugal. A total of 4716 animal samples were inoculated onto DERM agar, incubated at 25 °C for up to 4 weeks, and periodically examined macro- and micro-scopically to observe and evaluate fungal growth. Of these, 271 samples were removed due to contaminant fungi. Of the 568 positive cultures, the highest number were from the North (48.1%; 95% CI: 44.0-52.2%) and Centre (32.4%; 95% CI: 28.7-36.4%) regions. Microsporum canis was the most frequently isolated species (63.9%), followed by Trichophyton spp. (20.3%) and Nannizia gypsea (formerly Microsporum gypseum) (8.1%). Felines exhibited a higher frequency (17.4%) compared with dogs (9.1%) (p < 0.001). In dogs, the Yorkshire Terrier, West Highland White Terrier, Miniature Pinscher, Dalmatian and Miniature Schnauzer demonstrated a significant predisposition to dermatophytosis (p < 0.05). In cats, the Persian and Scottish Fold breeds were significantly predisposed (p < 0.05). No significant differences were found between sexes (p > 0.05). These findings underscore dermatophytosis as an increasing public health concern due to its zoonotic and contagious nature, providing comprehensive insights into the epidemiology of dermatophytosis in Portugal.

Serological diagnosis of Toxocara canis in mothers and their neonate referred to selected teaching hospitals, Tehran, Iran, 2020.

Toxocariasis is a common zoonotic parasitic disease, primarily caused by the larvae of Toxocara canis or T. cati. Humans become infected through accidental ingestion of infective eggs in soil contaminated with feces of infected animals, or by consuming undercooked meat from paratenic hosts harboring Toxocara larvae. This cross-sectional study aimed to evaluate the seroprevalence of Toxocara spp. in 250 mothers and their 250 neonates referred to selected teaching hospitals affiliated with Shahid Beheshti University of Medical Sciences, Tehran, Iran in 2020. Data were recorded using a specialized questionnaire, and serum and peripheral blood smears (for eosinophil count) were collected. Serum samples were tested using the ELISA serological test for IgG antibodies against Toxocara spp. The collected data were analyzed using IBM SPSS Statistics software version 21.0. A chi-square test was performed, and a P value less than 0.05 was considered statistically significant. Among the 500 participants, 24 cases (4.8%) tested positive for anti-Toxocara canis IgG antibodies. Seropositive cases included 6 mothers only (25%), 6 neonates only (25%), and 12 cases both mother and their neonate (50%). Statistical analysis showed a significant relationship between the presence of anti-Toxocara antibodies and hospital type and nationality (P value < 0.05). However, no significant relationship was found between the presence of antibodies and other demographic variables such as age, sex, eosinophilia, and history of contact with animals in mothers and their neonates (P value > 0.05). This study indicates a substantial rate Toxocara infection in mothers and neonates, necessitating appropriate preventive measures by the relevant authorities.

Hemoparasitism in grazing cattle and risk factors associated with husbandry management in an endemic area of Eastern Colombia

Vector-borne pathogens induce hemoparasitism in cattle causing substantial economic losses in tropical and subtropical areas. Infectious cattle actively contribute to maintaining the transmission cycle, and the presence of these animals must be associated with husbandry management and environmental changes. In the present study, we conducted a cross-sectional study sampling 1,000 bovines to identify infectious cattle diagnosed by a direct technique and employed a dichotomic questionnaire for association analyses, hierarchical clustering, and Principal Component Analysis (PCA). Overall prevalence with infectious cattle was 34.99%, where 97% of the farms had at least one infectious animal per genera, and the prevalence in properties ranged between 16.39 and 53.85%. Of these animals, 26.20% tested positive for Anaplasma sp., 8.40% for Babesia spp., and 1.30% for Trypanosome spp. The main co-infection showed 5% Anaplasma sp. – Trypanosome spp., followed by 4% Babesia spp. – Trypanosome spp. These bovines showed association with the use of the Jersey breed (OR = 2.016 C.I:1.188–3.419), selling animals for replacement (OR = 1.417 CI:1.022–1.965), participation in livestock exhibitions (OR = 2.009 CI:1.262–3.199), premises with burials (OR = 2,064 CR: 1.414–3.011), use of palm kernel (OR = 1.935 C.I:1.198–3.124), and the use of ivermectin (OR = 1.548 CI: 1.085–2.210) as a susceptibility. The hierarchical clustering revealed clusters among properties with different hemoparasite prevalence, with notable co-infections observed. The subsequent PCA identified that significant risk factors contributed to hemoparasitism positivity. We conclude that infectious cattle in the endemic area showed an association with husbandry management that permits the success of vector and maintenance of the enzootic or epizootic cycle in the herds.

Mouse hepatitis virus JHMV I protein is required for maximal virulence.

Betacoronaviruses encode a conserved accessory gene within the +1 open reading frame (ORF) of nucleocapsid called the internal N gene. This gene is referred to as "I" for mouse hepatitis virus (MHV), ORF9b for severe acute respiratory CoV (SARS-CoV) and SARS-CoV-2, and ORF8b for Middle East respiratory syndrome CoV (MERS-CoV). Previous studies have shown ORF8b and ORF9b have immunoevasive properties, while the only known information for MHV I is its localization within the virion of the hepatotropic/neurotropic A59 strain of MHV. Whether MHV I is an innate immune antagonist or has other functions has not been evaluated. In this report, we show that the I protein of the neurotropic JHM strain of MHV (JHMV) lacks a N terminal domain present in other MHV strains, has immunoevasive properties, and is a component of the virion. Genetic deletion of JHMV I (rJHMVIΔ57-137) resulted in a highly attenuated virus both in vitro and in vivo that displayed a post RNA replication/transcription defect that ultimately resulted in fewer infectious virions packaged compared with wild-type virus. This phenotype was only seen for rJHMVIΔ57-137, suggesting the structural changes predicted for A59 I altered its function, as genetic deletion of A59 I did not change viral replication or pathogenicity. Together, these data show that JHMV I both acts as a mild innate immune antagonist and aids in viral assembly and infectious virus production, and suggest that the internal N proteins from different betacoronaviruses have both common and virus strain-specific properties.IMPORTANCECoV accessory genes are largely studied in overexpression assays and have been identified as innate immune antagonists. However, functions identified after overexpression are often not confirmed in the infected animal host. Furthermore, some accessory proteins are components of the CoV virion, but their role in viral replication and release remains unclear. Here, we utilized reverse genetics to abrogate expression of a conserved CoV accessory gene, the internal N ("I") gene, of the neurotropic JHMV strain of MHV and found that loss of the I gene resulted in a post replication defect that reduced virion assembly and ultimately infectious virus production, while also increasing some inflammatory molecule expression. Thus, the JHMV I protein has roles in virion assembly that were previously underappreciated and in immunoevasion.

Veterinary perspectives on the urbanization of leishmaniosis in Morocco

BackgroundLeishmaniosis caused by Leishmania infantum, L. major and L. tropica is endemic in Morocco. Growing evidence of both human and canine Leishmania infections in urban centres has been reported. Since many forms of the disease are zoonotic, veterinarians play an important role in leishmaniosis control by intervening at the parasite host level. This study aimed to bring together One Health principles to connect canine and feline leishmaniosis epidemiology within urban centres of Morocco (Rabat and Fez) and assess the level of awareness of Moroccan veterinarians about facing this threat.MethodsA molecular survey was conducted for Leishmania DNA detection in canine (n = 155) and feline (n = 32) whole-blood samples. Three conventional polymerase chain reaction (PCR) protocols were implemented. The first PCR aimed at identifying infected animals by targeting Leishmania spp. kinetoplast minicircle DNA (kDNA). The second and third PCR targeted the Leishmania internal transcribed spacer region (ITS-1) and the Leishmania small subunit ribosomal RNA (SSUrRNA) gene, respectively, aiming at identification of the infecting species after Sanger sequencing-positive amplicons. Total immunoglobulin G (IgG) against Leishmania spp. was evaluated in 125 dogs by enzyme-linked immunosorbent assays (ELISA) using an in-house protocol, including three Leishmania-specific antigens (SPLA, rKDDR and LicTXNPx). Sera from 25 cats were screened for total IgG to Leishmania spp. by an indirect immunofluorescence antibody test (IFAT). An online questionnaire was presented to Moroccan veterinarians addressing their knowledge and practices towards animal leishmaniosis.ResultsOverall, 19.4% of the dogs tested positive for Leishmania kDNA and ITS-1 and sequencing revealed infection with L. infantum among PCR-positive dogs. These animals presented a wide range of ELISA seropositivity results (16.7%, 34.9% and 51.6%) according to the tested antigens (rKDDR, SPLA and LicTXNPx, respectively). Use of kDNA-PCR revealed 12.5% cats positive to Leishmania spp. otherwise found to be seronegative by IFAT.ConclusionsA considerable prevalence of infection was identified in dogs from urban centres of Morocco. Additionally, this is the first report of feline infection with Leishmania spp. in this country and in urban settings. Moroccan veterinarians are aware that animal leishmaniosis is endemic in Morocco, representing a public health threat, and are knowledgeable about canine leishmaniosis diagnosis and treatment.Graphical

Influenza A virus within-host evolution and positive selection in a densely sampled household cohort over three seasons.

While influenza A virus (IAV) antigenic drift has been documented globally, in experimental animal infections, and in immunocompromised hosts, positive selection has generally not been detected in acute infections. This is likely due to challenges in distinguishing selected rare mutations from sequencing error, a reliance on cross-sectional sampling, and/or the lack of formal tests of selection for individual sites. Here, we sequenced IAV populations from 346 serial, daily nasal swabs from 143 individuals collected over three influenza seasons in a household cohort. Viruses were sequenced in duplicate, and intrahost single nucleotide variants (iSNV) were identified at a 0.5% frequency threshold. Within-host populations were subject to purifying selection with >75% mutations present at <2% frequency. Children (0-5 years) had marginally higher within-host evolutionary rates than adolescents (6-18 years) and adults (>18 years, 4.4×10-6 vs. 9.42×10-7 and 3.45×10-6, p <0.001). Forty-five iSNV had evidence of parallel evolution, but were not overrepresented in HA and NA. Several increased from minority to consensus level, with strong linkage among iSNV across segments. A Wright Fisher Approximate Bayesian Computational model identified positive selection at 23/256 loci (9%) in A(H3N2) specimens and 19/176 loci (11%) in A(H1N1)pdm09 specimens, and these were infrequently found in circulation. Overall, we found that within-host IAV populations were subject to purifying selection and genetic drift, with only subtle differences across seasons, subtypes, and age strata. Positive selection was rare and inconsistently detected.

Correlates of disease severity in bluetongue as a model of acute arbovirus infection.

Most viral diseases display a variable clinical outcome due to differences in virus strain virulence and/or individual host susceptibility to infection. Understanding the biological mechanisms differentiating a viral infection displaying severe clinical manifestations from its milder forms can provide the intellectual framework toward therapies and early prognostic markers. This is especially true in arbovirus infections, where most clinical cases are present as mild febrile illness. Here, we used a naturally occurring vector-borne viral disease of ruminants, bluetongue, as an experimental system to uncover the fundamental mechanisms of virus-host interactions resulting in distinct clinical outcomes. As with most viral diseases, clinical symptoms in bluetongue can vary dramatically. We reproduced experimentally distinct clinical forms of bluetongue infection in sheep using three bluetongue virus (BTV) strains (BTV-1IT2006, BTV-1IT2013 and BTV-8FRA2017). Infected animals displayed clinical signs varying from clinically unapparent, to mild and severe disease. We collected and integrated clinical, haematological, virological, and histopathological data resulting in the analyses of 332 individual parameters from each infected and uninfected control animal. We subsequently used machine learning to select the key viral and host processes associated with disease pathogenesis. We identified and experimentally validated five different fundamental processes affecting the severity of bluetongue: (i) virus load and replication in target organs, (ii) modulation of the host type-I IFN response, (iii) pro-inflammatory responses, (iv) vascular damage, and (v) immunosuppression. Overall, we showed that an agnostic machine learning approach can be used to prioritise the different pathogenetic mechanisms affecting the disease outcome of an arbovirus infection.

The occurrence of Leptospira spp. serogroup Pomona infections in wild boars

Leptospirosis is a worldwide zoonosis caused by different pathogenic serovars of the genus Leptospira, with very complex aetiology and epidemiology. Recent publications suggest that wild boar (Sus scrofa) has an important role in the epizootiology of leptospirosis, representing a potential source of infection. Wild boar populations in urban environments are increasing, posing not only agricultural and ecosystem concerns, but also a public health concern. The aim of this study was to assess the prevalence, temporal patterns and risk factors of Pomona infection in wild boars from areas with pronounced human activity. A total of 159 wild boar serum samples were collected within a health monitoring project in Medvednica Nature Park over a period of six hunting seasons (2012–2016; 2018) and 25.2% samples tested positive. Seropositivity against seven different serogroups was observed, with the highest number of positive reactions to the serogroup Pomona. Annual variation in Pomona reactivity was also detected, and positive reactions were five times more common in females than in males. In most of studies, adults had a higher seroprevalence than younger categories; however, in this study,seroprevalence against Pomona was higher in piglets and subadults. The results indicate that wild boar has the potential of maintaining serovars from the serogroup Pomona in certain natural foci. Due to the expansion of wild boar populations and their intrusion into urban areas, they should be considered a potentially important source of infection for humans and domestic animals.

Bat Rabies in the Americas: Is Myotis the Main Ancestral Spreader?

The rabies virus (RABV) is the exclusive lyssavirus affecting both wild and domestic mammalian hosts in the Americas, including humans. Additionally, the Americas stand out as the sole region where bat rabies occurs. While carnivore rabies is being increasingly managed across the region, bats are emerging as significant reservoirs of RABV infection for humans and domestic animals. Knowledge of the bat species maintaining rabies and comprehending cross-species transmission (CST) and host shift processes are pivotal for directing surveillance as well as ecological research involving wildlife reservoir hosts. Prior research indicates that bat RABV CST is influenced by host genetic similarity and geographic overlap, reflecting host adaptation. In this study, we compiled and analyzed a comprehensive nucleoprotein gene dataset representing bat-borne RABV diversity in Argentina and the broader Americas using Bayesian phylogenetics. We examined the association between host genus and geography, finding both factors shaping the global phylogenetic structure. Utilizing a phylogeographic approach, we inferred CST and identified key bat hosts driving transmission. Consistent with CST determinants, we observed monophyletic/paraphyletic clustering of most bat genera in the RABV phylogeny, with stronger CST evidence between host genera of the same family. We further discuss Myotis as a potential ancestral spreader of much of RABV diversity.

Recent Advances in the Development of Metal/Metal Oxide Nanoparticle and Antibiotic Conjugates (MNP-Antibiotics) to Address Antibiotic Resistance: Review and Perspective.

As per the World Health Organization (WHO), antimicrobial resistance (AMR) is a natural phenomenon whereby microbes develop or acquire genes that render them resistant. The rapid emergence and spread of this phenomenon can be attributed to human activity specifically, the improper and excessive use of antimicrobials for the treatment, prevention, or control of infections in humans, animals, and plants. As a result of this factor, many antibiotics have reduced effectiveness against microbes or may not work fully. Thus, there is a pressing need for the development of new antimicrobial agents in order to counteract antimicrobial resistance. Metallic nanoparticles (MNPs) are well known for their broad antimicrobial properties. Consequently, the use of MNPs with current antibiotics holds significant implications. MNPs, including silver nanoparticles (AgNPS), zinc oxide nanoparticles (ZnONPs), copper nanoparticles (CuNPs), and gold nanoparticles (AuNPs), have been extensively studied in conjunction with antibiotics. However, their mechanism of action is still not completely understood. The interaction between these MNPs and antibiotics can be either synergistic, additive, or antagonistic. The synergistic effect is crucial as it represents the desired outcome that researchers aim for and can be advantageous for the advancement of new antimicrobial agents. This article provides a concise and academic description of the recent advancements in MNP and antibiotic conjugates, including their mechanism of action. It also highlights their possible use in the biomedical field and major challenges associated with the use of MNP-antibiotic conjugates in clinical practice.

Novel Pestiviruses Detected in Cattle Interfere with Bovine Viral Diarrhea Virus Diagnostics.

Since the start of the mandatory nationwide bovine viral diarrhea (BVD) eradication program in Germany in 2011, the number of persistently infected (PI) animals has decreased considerably, resulting in a continuous decrease in seroprevalence. The increasingly BVD-naive cattle population could facilitate spillover infections with non-BVDV ruminant pestiviruses. Here, we report two cases in which novel pestiviruses were isolated from cattle; in both cases, the whole genome sequence showed the highest level of identity to strain "Pestivirus reindeer-1". Both novel viruses gave positive results in BVDV diagnostic test systems, confirming that cross-reactivity is an important issue in pestivirus diagnostics. In the first case, the pestivirus was probably transmitted from sheep kept with the affected cattle, suggesting that the co-housing of small ruminants and cattle is a risk factor. The source of infection could not be determined in the second case. The occurrence of these two cases in independent cattle holdings within a relatively short time frame suggests that it would be useful to determine the presence of pestiviruses in small ruminants or even wild ruminants to better assess risk factors, especially for BVDV-free populations.